Team:Paris/September 1

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(Construction of pFlhB - mRFP Tripart (LVA+))
 
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{{Paris/Calendar_Links|August 31|September 2}}
{{Paris/Calendar_Links|August 31|September 2}}
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=Cloning of EnvZ*=
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==Transformation results of L165 (EnvZ*-pSB1A2) from August 29th==
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{|border="1" style="text-align: center"
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|'''sample'''
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|'''number of colonies'''
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|-
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|'''positive''' control pUC19
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|many colonies
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|-
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|transformation '''negative''' control
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|0
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|-
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|ligation '''without insert'''
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|0
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|-
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|'''3:1''' insert:vector ratio
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|5
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|-
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|'''4:1''' insert / vector ratio
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|5
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|-
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|}
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=Construction of rbs-FlhDC-mRFP-LVA-tripart (L172)=
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Aim : Construction of ''' "pFlhB-RBS-mRFP-dbl ter" (pFlhB-I732078)''' [[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
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<br>==> it appears a mistake of name digestion, it explains the realisation of a construction not wanted
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==Digestion==
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==='''Measurement of the concentration of D187 purified'''===
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[[Team:Paris/Notebook/Protocols#Concentration_of_the_Miniprep | Protocol (it's same that for Miniprep)]]
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{|border="1" style="text-align: center"
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|'''Digestion Name'''
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|'''Concentration (µg/mL)'''
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|'''Ratio 260/280'''
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|-
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|D187
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|16
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|1.31
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|-
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|D186
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|24
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|1.25
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|}
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==Ligation==
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[[Team:Paris/Notebook/Protocols#Ligation |Protocol]]
{|border="1" style="text-align: center"
{|border="1" style="text-align: center"
-
|
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|'''Ligation Name'''
-
|'''DNA concentration'''
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|'''Vector Name'''
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|'''A260 / A2801'''
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|'''Volume Vector (µL)'''
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|'''Insert'''
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|'''Volume Insert (µL)'''
|-
|-
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|D159 (EnvZ*)
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|L168
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|9 µg/mL
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|D187
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|1,59
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|2.5
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|D186
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|1
|-
|-
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|D116 (pSB1A2)
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|Control L168
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|13 µg/mL
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|D166
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|1,29
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|2.5
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| -
|}
|}

Latest revision as of 14:49, 13 September 2008

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Contents

Cloning of EnvZ*

Transformation results of L165 (EnvZ*-pSB1A2) from August 29th

sample number of colonies
positive control pUC19 many colonies
transformation negative control 0
ligation without insert 0
3:1 insert:vector ratio 5
4:1 insert / vector ratio 5


Construction of rbs-FlhDC-mRFP-LVA-tripart (L172)

Aim : Construction of "pFlhB-RBS-mRFP-dbl ter" (pFlhB-I732078) Part icon regulatory.pngPart icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
==> it appears a mistake of name digestion, it explains the realisation of a construction not wanted

Digestion

Measurement of the concentration of D187 purified

Protocol (it's same that for Miniprep)

Digestion Name Concentration (µg/mL) Ratio 260/280
D187 16 1.31
D186 24 1.25

Ligation

Protocol

Ligation Name Vector Name Volume Vector (µL) Insert Volume Insert (µL)
L168 D187 2.5 D186 1
Control L168 D166 2.5 -